Kronborg I J, Knopf P M
Ann Acad Med Singap. 1980 Apr;9(2):176-8.
A method has been developed to measure the in vitro production of immunoglobulin (Ig) by liver biopsy specimens. Five to 30 mg of liver tissue was cultured for 24 h in Dulbecco's modified Eagle's medium/10% foetal calf serum (FCS) containing radiolabelled leucine (L-[4,5-3H] leucine). The culture medium was collected, centrifuged and the supernatant dialysed to remove labelled leucine. The residual radioactivity was a measure of newly synthesized 3H-labelled proteins released into the medium. The quantity of IgG was determined by immunoprecipitation with monospecific antisera to IgG heavy chains. The presence of IgG in the supernatant was confirmed by chromatography on protein-A Sepharose column. In 6 biopsies without evidence of active inflammation (4 normal and 2 fatty liver by histological criteria) less than 1% of the protein synthesized was IgG. In contrast in the presence of active inflammation in 4 cases of alcoholic hepatitis the IgG percentage ranged from 2 to 6%. Maximal levels of IgG production were detected in 3 cases of chronic active hepatitis (CAH) and ranged from 5 to 30%. The increased Ig synthesis by the liver in alcoholic hepatitis and CAH is presumed to be an index of the intrahepatic host response and may have important implications for mechanisms of liver damage in these diseases.
已开发出一种方法来测量肝活检标本中免疫球蛋白(Ig)的体外产生量。将5至30毫克肝组织在含有放射性标记亮氨酸(L-[4,5-³H]亮氨酸)的杜尔贝科改良伊格尔培养基/10%胎牛血清(FCS)中培养24小时。收集培养基,离心,然后将上清液透析以去除标记的亮氨酸。残留放射性是释放到培养基中的新合成的³H标记蛋白质的量度。通过用针对IgG重链的单特异性抗血清进行免疫沉淀来测定IgG的量。通过在蛋白A琼脂糖柱上进行色谱分析来确认上清液中IgG的存在。在6例无活动性炎症证据的活检标本中(根据组织学标准,4例正常,2例脂肪肝),合成的蛋白质中IgG含量不到1%。相比之下,在4例酒精性肝炎伴有活动性炎症的情况下,IgG百分比范围为2%至6%。在3例慢性活动性肝炎(CAH)中检测到IgG产生的最高水平,范围为5%至30%。酒精性肝炎和CAH中肝脏Ig合成增加被认为是肝内宿主反应的一个指标,可能对这些疾病中肝损伤的机制具有重要意义。